Fatty acids, C5-10, esters with pentaerythritol

Short-term toxicity to aquatic invertebrates

Link to relevant study record(s)

Reference

Reference 1

Endpoint:

short-term toxicity to aquatic invertebrates

Type of information:

experimental study

Adequacy of study:

disregarded due to major methodological deficiencies

Study period:

03 February to 03 April 2000

Reliability:

3 (not reliable)

Rationale for reliability incl. deficiencies:

other: Inconsistent result (test substance film at the highest concentration, number of daphnids trapped at the surface at each concentration)

Qualifier:

according to guideline

Guideline:

ISO 6341 (Water quality - Determination of the Inhibition of the Mobility of Daphnia magna Straus (Cladocera, Crustacea))

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)

Qualifier:

equivalent or similar to guideline

Guideline:

EU Method C.2 (Acute Toxicity for Daphnia)

GLP compliance:

yes

Analytical monitoring:

no

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: Water accommodated fractions (WAFs); weighed amounts of test substance expressed as mg/l were mixed with test medium, treated with
ultrasonic waves and stirred for ca. 48 hours in a closed vessel in the dark; the dispersions were allowed to settle overnight until a stable separation
of liquid phases was reached.

Test organisms (species):

Daphnia magna

Details on test organisms:

TEST ORGANISM
Common name: Water flea
- Strain: Straus
- Age at study initiation (mean and range, SD): < 24 hours

HOUSING
- Housing conditions: Maximum age of the cultures was 4 weeks, renewal of the cultures was after 7 days of cultivation half of the medium twice a
week
- Type and amount of food: A suspension of fresh water algae
- Feeding frequency: Daily

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

48 h

Test temperature:

20.5-21.8 °C (control)

pH:

7.8-8.0

Dissolved oxygen:

Start (t=0 h): 9.2-9.7
End (t=48 h): 8.8-9.2

Nominal and measured concentrations:

Water Accommodated Fractions (WAFs) prepared at: 10, 18, 32, 56 and 100 mg/L

Details on test conditions:

TEST SYSTEM
- Test vessel: 100 ml, all-glass
- Material, size, headspace, fill volume: 80 ml fill volume
- Aeration: none
- No. of organisms per vessel: 10
- No. of vessels per concentration (replicates): 2
- No. of vessels per control (replicates): 2

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: ISO, prepared in milli-RO water
- Culture medium different from test medium: yes, culture medium was medium M7 as prescribed by Elendt-Schneider (Elendt, 1990), trace elements, macro nutrients and vitamins are added to freshly prepared ISO-medium
- Intervals of water quality measurement: pH and dissolved oxygen at the beginning and at the end of the test

OTHER TEST CONDITIONS
- Photoperiod: 16 hours light daily

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) : immobility at 24 and 48 hours

TEST CONCENTRATIONS
- Range finding study
- Test concentrations: WAFs prepared at 10 and 100 mg/l and dilutions containing 1 and 10% of the 10 mg/l WAF
- Results used to determine the conditions for the definitive study: All daphnids exposed to a loading rate of 100 mg/l (WAF) are immobilised at the end of the test period. No immobility was observed in the lower test concentrations.

Reference substance (positive control):

yes

Remarks:

potassium dichromate

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

78 mg/L

Nominal / measured:

nominal

Basis for effect:

mobility

Remarks on result:

other: 95% confidence interval between 68 and 98 mg/L

Duration:

48 h

Dose descriptor:

NOEC

Effect conc.:

32 mg/L

Nominal / measured:

nominal

Basis for effect:

mobility

Details on results:

- Behavioural abnormalities: A number of daphnids trapped at the surface at each concentration. At a test concentration of 18 mg/L daphnids
stucked to the bottom of the test vessel. Before recording of immobility the daphnids were reimmersed in the test solution.
- Mortality of control: none
- Any observations (e.g. precipitation) that might cause a difference between measured and nominal values: At 100 mg/L (WAF) a test substance film was observed after 24 hours of exposure.
- Effect concentrations exceeding solubility of substance in test medium: yes

Results with reference substance (positive control):

- Results with reference substance valid? yes
- EC50/LC50: EC50 (48h) of potassium dichromate was 0.45 mg/L (95% fiducial limits of 0.41 - 0.53 mg/L)

At each test concentration daphnids trapped at the surface of the treated test solutions.

At 100 mg/L test concentration the daphnids showed an immobilisation of > 50% (final test: 75% (mean), range finding test: 100%) after 48h exposure. A test substance film was observed after 24 hours of exposure at this highest concentration. The EC50 was stated as 78 mg/l (nominal). Because of the test substance film at the highest concentration and the trapped daphnids at the surface at each concentration, the test is not reliable. The effect at 100 mg/L could result in physical effects caused in undissolved substance.

Table 1: Acute immobilisation of Daphnia magna after 24 and 48 hours

(between brackets: number of daphnids observed trapped at the surface)

Test substance (WAF at x mg/l)	Vessel number	Number Daphnia exposed	Immobilisation at 24h		Immobilisation at 48 h
			number	%	number	%
Control	A B	10 10	0 0	0 0	0 0	0 0
10	A B	10 10	0 (7) 0 (3)	0 0	1 (1) 0 (2)	10 0
18	A B	10 10	3 (10) 0 (6)	30 0	0 1 (2)	0 10
32	A B	10 10	0 (10) 0 (10)	0 0	0 0 (1)	0 0
56	A B	10 10	0 (6) 0 (5)	0 0	3 (1) 0	30 0
100	A B	10 10	3 (10) 2 (9)	30 20	7 8	70 80

 

Reference 2

Endpoint:

short-term toxicity to aquatic invertebrates

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: see 'Remark'

Remarks:

GLP - Guideline study, tested with the source substance Fatty acids, C5-9 tetraesters with pentaerythritol (CAS 67762-53-2). According to the ECHA guidance document “Practical guide 6: How to report read-across and categories (March 2010)”, the reliability was changed from RL1 to RL2 to reflect the fact that this study was conducted on a read-across substance.

Qualifier:

according to guideline

Guideline:

OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)

Qualifier:

according to guideline

Guideline:

EU Method C.2 (Acute Toxicity for Daphnia)

Qualifier:

according to guideline

Guideline:

ISO 6341 (Water quality - Determination of the Inhibition of the Mobility of Daphnia magna Straus (Cladocera, Crustacea))

Qualifier:

according to guideline

Guideline:

other: OECD series on testing and assessment 23, 2000

Principles of method if other than guideline:

Test conducted by preparing WAFs due to low water solubility of the test substance.

GLP compliance:

yes

Analytical monitoring:

yes

Details on sampling:

- Concentrations: During the final test singular samples for possible analysis were taken from all test concentrations and the control at test start and test end
- Sampling method: 2 mL from the approximate centre of the test vessels
- Sample storage conditions before analysis: Samples were stored in a freezer until analysis

At the end of the exposure period, the replicates were pooled at each concentration before sampling. Additionally, reserve samples of 2 mL were taken for possible analysis. If not used, these samples were stored in a freezer for a maximum of three months after delivery of the draft report, pending on the decision of the sponsor for additional analysis

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: Preparation of test solutions started with a loading rate of 100 mg/L applying 2 days of magnetic stirring to ensure maximum dissolution in test medium. The resulting dispersion was left to settle for 1 hour (range-finding test) or 2 hours (final test) were after the Water Accomodated Fraction (WAF) was collected by siphoning for use as highest test concentration. The lower test concentrations were prepared by subsequent dilutions in test medium.
- Evidence of undissolved material (e.g. precipitate, surface film, etc): The final test solutions were all clear and colourless.

Test organisms (species):

Daphnia magna

Details on test organisms:

TEST ORGANISM
- Common name: water flea
- Source: In-house laboratory culture with a known history. Daphnids originated from a healthy stock, 2nd to 5th brood, showing no signs of stress such as mortality >20%, presence of males, ephippia or discoloured animals and there was no delay in the production of the first brood
- Age at study initiation: <24 hours, from parental daphnids of more than two weeks old
- Method of breeding: Start of each batch with newborn daphnids, i.e. less than 3 days old, by placing about 250 of them into 5 litres of medium in an all-glass culture vessel. Maximum age of the cultures is 4 weeks. Renewal of the cultures: after 7 days of cultivation half of the medium twice a week. Temperature of medium (M7, Elendt): 18 - 22 °C. Daphnids werde fed daily with a suspension of fresh water algae.
- Feeding during test: no

Test type:

static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

48 h

Hardness:

180 mg/L expressed as CaCO3

Test temperature:

19.8 °C - 20.4 °C

pH:

7.9 - 8.0 (final test)

Dissolved oxygen:

8.7 - 8.9 mg/L (final test)

Nominal and measured concentrations:

combined limit/range finding test: control, 0.1, 1.0, 10 and 100% of the WAF prepared at a loading rate of 100 mg/L (nominal)
final test: control, 4.6, 10, 22, 46 and 100% of the WAF prepared at a loading rate of 100 mg/L (nominal)
final test: control, 0.045, 0.18, 0.47, 0.63, and 1.3 mg/L (measured)

Details on test conditions:

TEST SYSTEM
- Test vessel:
- Type: open
- Material, size, fill volume: glass vessels with a barrier net just below the surface to prevent Daphnids to swim to the surface in order to avoid contact to any undissolved test material, 90-mL, 70 mL fill volume
- Aeration: no
- No. of organisms per vessel: 5 (combined limit/range finding test and final test)
- No. of vessels per concentration (replicates): 4 (final test), 4 replicates at 100% WAF, 2 at 0.1, 1.0, and 10% (combined limit/range finding test)
- No. of vessels per control (replicates): 4 (combined limit/range finding test and final test)

TEST MEDIUM / WATER PARAMETERS
- Culture medium different from test medium: no; M7 medium after Elendt (Elendt, B.-P., 1990: Selenium deficiency in Crustacea. An ultrastructural approach to antennal damage in Daphnia magna Straus. Protoplasma 154, 25-33)


OTHER TEST CONDITIONS
- Adjustment of pH: no
- Photoperiod: 16 hours light, 8 hours dark

EFFECT PARAMETERS MEASURED (with observation intervals if applicable): immobility at 24 hours and 48 hours

TEST CONCENTRATIONS
- Spacing factor for test concentrations: 10 in combined limit/range finding test, approx. 2 in final test
- Range finding study
- Test concentrations: 0.1, 1.0, 10 and 100% of the WAF
- Results used to determine the conditions for the definitive study: The two highest test groups contained a floating layer from 24 hours onwards causing entrapment of a significant number of organisms. Additional microscopic evaluation performed after 24 hours showed that bodies of the daphnids in the highest test group were all covered with test article. Hence, at least part if not all of the effects were of physical nature and caused by exposure to undissolved test material. It was concluded that the test set-up for the final study should include a barrier to prevent organisms getting stuck in the floating layer.

Reference substance (positive control):

yes

Remarks:

potassium dichromate

Key result

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

> 1.3 mg/L

Nominal / measured:

meas. (geom. mean)

Conc. based on:

test mat.

Basis for effect:

mobility

Key result

Duration:

48 h

Dose descriptor:

EL50

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

other: test mat. WAF

Basis for effect:

mobility

Key result

Duration:

48 h

Dose descriptor:

NOEC

Effect conc.:

0.63 mg/L

Nominal / measured:

meas. (geom. mean)

Conc. based on:

test mat.

Basis for effect:

mobility

Key result

Duration:

48 h

Dose descriptor:

NOELR

Effect conc.:

46 other: % of a WSF prepared at a loading rate of 100 mg/L

Nominal / measured:

nominal

Conc. based on:

other: test mat. WAF

Basis for effect:

mobility

Details on results:

- Effect concentrations exceeding solubility of substance in test medium: yes

Limit/range finding test: Immobility at 10 and 100% WAF were at respectively 50 and 95% at the end of the test. It should however be recognized that the two highest test groups contained a floating layer from 24 hours onwards causing entrapment of a significant number of organisms. Additional microscopic evaluation performed after 24 hours showed that bodies of the daphnids in the highest test group were all covered with test article. Hence, at least part if not all of the effects were of physical nature and caused by exposure to undissolved test material.

4.8 Samples taken from 1.0 and 100% of the WAF prepared at 100 mg/L were analysed. The initial concentration in the undiluted WAF was 2.2 mg/L. This concentration decreased to 0.22 mg/L after 48 hours. In samples taken from 1.0% WAF, the measured concentration decreased from 0.17 mg/L at the start to a concentration below the limit of detection during the test period. Concentrations in both solutions decreased significantly during the test period, which was expected to be related to the extremely low water solubility (< 6 µg/L).

Analytical results final test: Analysis of the samples taken at the start of the final test showed measured concentrations of 0.27, 0.85, 1.9, 3.6 and 7.0 mg/L in solutions representing 4.6, 10, 22, 46 and 100% of the WSF, respectively. The concentrations decreased during the test period to approximately 3 - 6% of initial. However, measured concentrations in all solutions at the end of the study still exceeded the water solubility that was determined to be less than 6 µg/L. Based on these results, the average exposure concentrations were calculated to correspond with 0.045, 0.18, 0.47, 0.63 and 1.3 mg/L.

Contrary to the combined limit/range-finding test, no daphnids became trapped at the surface despite the fact that initial exposure concentrations in the undiluted WSF were even higher than those measured in the combined limit/range-finding test. This was the result of the use of a physical barrier in combination with placing the light source underneath the vessels. As a consequence, only very few daphnia were immobilised in comparison to the same solutions tested in the combined limit/range-finding test. This confirmed that effects in the range-finding were of physical instead of toxic nature.

Results with reference substance (positive control):

- Results with reference substance valid? yes
- EC50 = 0.3 mg/L (95% CL: 0.28 - 0.35 mg/L

Table: Incidence of immobility in the combined limit/range-finding test

Test group Fatty acids, C5-9, tetraesters with pentaerythritol (% WSF*)	Vessel number	Number Daphnia exposed	Response at 24 h		Response at 48 h
			number	Total %	number	Total %
Control	A B C D	5 5 5 5	0 0 0 0	0	0 0 0 0	0
0.1	A B	5 5	0 0	0	0 0	0
1.0	A B	5 5	0 0	0	0 0	0
10	A B	5  3^1	0 (4) 0 (2)	0	2 (1) 2	50
100	A B C D	5 5 5  6^1	3 (5) 4 (5) 5 (5) 5 (5)	81	5 4 (1) 5 6	95

* = Test groups represent percentages of a WAF prepared at 100 mg/L

1 = Incorrect number of organisms exposed

( ) = number of Daphnia observed trapped at the surface of test solutions. These organisms were reimmersed into the respective solutions before recording of mobility

Reference 3

Endpoint:

short-term toxicity to aquatic invertebrates

Type of information:

experimental study

Adequacy of study:

key study

Study period:

8 Nov - 30 Nov 2005

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: see 'Remark'

Remarks:

GLP - Guideline study, tested with the source substance Decanoic acid, mixed esters with heptanoic acid, octanoic acid, pentaerythritol and valeric acid (CAS 71010-76-9). According to the ECHA guidance document “Practical guide 6: How to report read-across and categories (March 2010)”, the reliability was changed from RL1 to RL2 to reflect the fact that this study was conducted on a read-across substance.

Qualifier:

according to guideline

Guideline:

OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)

Deviations:

no

GLP compliance:

yes

Analytical monitoring:

yes

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION
- Method: Individual treatments were prepared by adding the appropriate amount of test substance to 2 L of laboratory dilution water in glass aspirator bottles. The test substance was added to the water in the aspirator bottles using stainless steel and glass syringes. The syringes were weighed before and after adding the test substance to determine the actual loading rate. The mixing vessels were closed with foil-covered neoprene stoppers. The mixtures were stirred using a 6.7% (of the static liquid depth) vortex for 23.5 h on magnetic stir plates with Teflon-coated stribars at room temperature. As stirring initiated and after stirring all treatments appeared clear and colourless with the test substance floating at the surface. The mixtures were allowed to settle and equilibrate to test temperature in a waterbath at 17.8 °C for one hour without stirring before removing the aqueous portions (WAFs) through the outlet at the bottom of the vessel for testing.

Test organisms (species):

Daphnia magna

Details on test organisms:

TEST ORGANISM
- Common name: water flea
- Source: in-house culture; original culture was supplied by Aquatic Biosystems, Fort Collins, USA on April 11, 2002
- Age at study initiation: < 24 h
- Feeding during test: no

ACCLIMATION
- Acclimation period: Day 0 cultures were started daily using 8 < 24 h old neonates from culture beakers between 12 and 18 days old, exhibiting <20% adult mortality
- Acclimation conditions. same as test
- Type and amount of food: Pseudokirchneriella subcapitata (~ 4.5E5 cells/mL) and 4 mL of a yeast/salmon starter/ Microfeast mixture per 800 mL
- Feeding frequency: daily (five days per week at a minimum)
- Health during acclimation: The pool of neonates selected for the test had ≤ 10% daily mortality on the experimental start day.

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

48 h

Hardness:

150 mg/L as CaCO3

Test temperature:

20.1 - 20.9

pH:

7.9 - 8.1

Dissolved oxygen:

7.3 - 8.3 mg/L

Nominal and measured concentrations:

Nominal: 62, 132, 251, 503 and 1072 mg/L loading rate WAF

Details on test conditions:

TEST SYSTEM
- Test vessel: Erlenmeyer flask
- Type: closed with ground glass stoppers
- Material: glass, size: 125 mL, fill volume: ~140 mL (no headspace)
- No. of organisms per vessel: 5
- No. of vessels per concentration: 4
- No. of vessels per control: 4

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: The dilution water was prepared from UV-sterilized, deionized well water that was treated and distributed throughout the testing facility via PVC and stainless steel pipes.
- Ca/mg ratio: 1.2:1
- Alkalinity: 96 mg/L as CaCO3
- Conductivity: 500 µmhos
- Culture medium different from test medium: no
- Intervals of water quality measurement: Water quality measurements (pH, dissolved oxygen, temperature) were performed on a sub-sample of the WAFs from each treatment and control on day 0 and on a composite of the replicates in each treatment at termination.

OTHER TEST CONDITIONS
- Photoperiod: 16 h light/ 8 h dark
- Light intensity: Daylight intensity ranged from ~184 - 233 lux during full daylight periods of the study.

EFFECT PARAMETERS MEASURED: Immobilisation was recorded after 24 and 48 h test duration. In addition, observations for normal or abnormal behaviour/ appearance were made.

TEST CONCENTRATIONS
- Spacing factor for test concentrations: 10
- Range finding study: The toxicity of the "water-accomnodated fraction" of the test substance was tested at different loading levels. The WAFs were prepared by adding the appropriate amount of test substance to dilution water in glass aspirator bottles. The vessels were closed and mixed on magnetic stir plates with Teflon coated stirbars for 23 h and 50 minutes. The treatments were allowed to settle for 1 h and five minutes after mixing. Two replicates at each loading level were prepared containing 5 organisms. This study was not conducted according to GLP.
- Test concentrations: 10, 100 and 1000 mg/L loading rate WAFs
- Results used to determine the conditions for the definitive study: No mortality at any loading level was recorded during a exposure time of 48 hours.

Reference substance (positive control):

no

Key result

Duration:

48 h

Dose descriptor:

EL50

Effect conc.:

> 1 072 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Remarks:

(WAF)

Basis for effect:

mobility

Details on results:

- Behavioural abnormalities: none

TOC analysis confirmed the low water solubility of the test substance in water. The total carbon content of the treatment is only slightly higher than that of the control. Thus, the results of the TOC analysis can not be used for the determination of an effect concentration. Thus, the effect concentration is related to the nominal concentration.

Table 1: TOC analysis of the test substance in the test medium

TOC [mg]
Loading rate	0 h	48 h
Control	0.4127	0.4102
62	0.4716	0.4246
132	0.4349	0.4330
251	0.3856	0.3966
503	0.4420	0.4097
1072	0.4415	0.4079

Reference 4

Endpoint:

short-term toxicity to aquatic invertebrates

Type of information:

experimental study

Adequacy of study:

key study

Study period:

17 - 19 Jan 2007

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Guideline study with acceptable restrictions

Qualifier:

according to guideline

Guideline:

OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)

Qualifier:

according to guideline

Guideline:

EU Method C.2 (Acute Toxicity for Daphnia)

GLP compliance:

no

Analytical monitoring:

no

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: Water Accommodated Fraction (WAF): This WAF was prepared by moderately stirring 100 mg test substance per liter of medium overnight
followed by filtration (MILLIPORE AP15 glass fiber filter).

Test organisms (species):

Daphnia magna

Details on test organisms:

TEST ORGANISM
- Common name: Water flea
- Strain: Straus
- Length at study initiation (length definition, mean, range and SD): < 2mm
- Method of breeding: Parental and young Daphnia were held in 40L glass aquaria.

BREEDING
- Breeding conditions (same as test or not): Reconstituted water (ISO 6341) was the same as test, prepared with distilled water. The medium in the
breeding system was continuously aerated in comparison to the test.
- Type and amount of food: 15 g sera micropan (sera GmbH, Heinsberg) homogenized in 1L deionized water
- Feeding frequency: Once a day, except weekends

ACCLIMATION
- Acclimation period: Daphnia, which had been selected in size, were acclimatized to the reconstituted water for 4 hours before introduction into
the test media.

Test type:

static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

48 h

Test temperature:

20.0 ± 0.5 °C

pH:

control: 7.3 at 0h, 7.7 at 48h
100 mg/l: 7.5 at 0h, 7.7 at 48h

Dissolved oxygen:

control: 6.2 mg/l at 0h, 6.0 mg/l at 48h
100 mg/l: 6.0 mg/l at 0h, 5.9 mg/l at 48h

Nominal and measured concentrations:

nominal: 100 mg/L (WAF)

Details on test conditions:

TEST SYSTEM
- Test vessel:
- Material, size, headspace, fill volume: 100 mL flasks, all-glass, with 50 mL of test medium
- Aeration: no
- No. of organisms per vessel: 20
- No. of vessels per concentration (replicates): 2
- No. of vessels per control (replicates): 2

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: Reconstituted water (ISO 6341), prepared with distilled water. The ratio of Ca:Mg ions is 4:1 and of Na:K ions is 10:1.
- Alkalinity: 0.8 mmol/L
- Ca/mg ratio: 4:1
- Culture medium different from test medium: No, but test medium was not aerated as the culture medium.
- Intervals of water quality measurement: Temperature determined in a control flask at the start and at the end of the test. Oxygen and pH were
determined in all vessels prior to the addition of Daphnia to the test medium and at the end of the test.

OTHER TEST CONDITIONS
- Photoperiod: 16 h a day, supplied by overhead white fluorescent tubes.

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) : Observations of immobile Daphnia were made after 24 and 48 h of
exposure

Reference substance (positive control):

no

Key result

Duration:

48 h

Dose descriptor:

EL50

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Key result

Duration:

48 h

Dose descriptor:

EL0

Effect conc.:

>= 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

The test substance show no acute toxicity for daphnids under test conditions.

Table 1: Immobilisation of Daphnia magna after 24 and 48 hours

Nominal concentration (mg/l)	Code	Number of individuals per test vessel	Immobile Daphnia (No./vessel) 24h	Immobile Daphnia (No./vessel) 48h	% immobile Daphnia after 24h of exposure	% immobile Daphnia after 48h of exposure
Control 0	A B C	20 20 20	0 1 0	0 1 0	0 5 0	0 5 0
100	A B	20 20	0 1	1 1	0 5	5 5

 

Reference 5

Endpoint:

short-term toxicity to aquatic invertebrates

Type of information:

experimental study

Adequacy of study:

key study

Study period:

21 Jul - 24 Aug 1995

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: see 'Remark'

Remarks:

GLP - Guideline study, tested with the source substance Mixed esters of pentaerythritol, technical grade, with isooctanoic and C8/C10 fatty acids (CAS 189200-42-8). According to the ECHA guidance document “Practical guide 6: How to report read-across and categories (March 2010)”, the reliability was changed from RL1 to RL2 to reflect the fact that this study was conducted on a read-across substance.

Qualifier:

according to guideline

Guideline:

OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)

GLP compliance:

yes

Analytical monitoring:

yes

Details on sampling:

- Concentrations: control, 1000 mg/L
- Sampling method: Samples were removed from the test solutions on day 0 and at termination.

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION
- Method: Individual treatment solutions (WAF) were prepared by adding the appropriate amount of the test substance to 1 L of dilution water in 1 L size glass aspirator bottles. The vessels were closed with cotton-gauze stoppers to minimize evaporation and/or volatilazition. The solutions were mixed at <10% vortex on magnetic stirplates with Teflon® coated stirbars for approximately 24 hours. All treatment levels appeared clear with test substance floating on the surface. After mixing, the solutions were allowed to settle for approximately one hour before the WAFs were removed through the outlet at the bottom of the aspirator bottle.
- Evidence of undissolved material: No test substance insolubility was observed in any of the test chambers.

Test organisms (species):

Daphnia magna

Details on test organisms:

TEST ORGANISM
- Common name: water flea
- Source: in-house culture
- Age at study initiation: < 24 hold, taken from 16 d old parents
- Method of breeding: Daphnids are kept in 1 L glass culture vessels with dilution water (BW3 at 20 ± 2 °C) supplemented with vitamin B12 and selenium.
- Feeding during test: none

ACCLIMATION
- Acclimation period: not specified
- Acclimation conditions: same as test
- Type and amount of food: Selenastrum capricornutum and a 6 µL/mL yeast/ salmon starter/ cereal leaves mixture
- Feeding frequency: daily

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

48 h

Hardness:

190 mg/L as CaCO3

Test temperature:

21.2 - 22.0 °C

pH:

7.6 - 8.0

Dissolved oxygen:

7.0 - 8.3 mg/L

Nominal and measured concentrations:

Nominal: 62.5, 125, 250, 500 and 1000 mg/L

Details on test conditions:

TEST SYSTEM
- Test vessel: Erlenmeyer flask sealed with ground glass stoppers
- Material: glass; Size: 125 mL; Fill volume: 140 mL of solution
- No. of organisms per vessel: 5
- No. of vessels per concentration: 4
- No. of vessels per control: 4

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: Laboratory dilution water, a mixture of carbon filtered well water and water dialyzed by reverse osmosis supplemented with 1 µg/L vitamin B12 and 2 µg/L selenium, aged and aerated ≥ 24 h prior to introduction of the test substance
- Total organic carbon: 1.528 ppm
- Alkalinity: 57 mg/L as CaCO3
- Conductivity: 370 µmhos
- Culture medium different from test medium: no
- Intervals of water quality measurement: Water quality measurements (pH, dissolved oxygen and temperature) were performed on a subsample of each treatment on Day 0 and a composite of the replicates at termination.

OTHER TEST CONDITIONS
- Photoperiod: 16 h light : 8 h dark with a gradual intensity conversion between periods
- Light intensity: 640.56 - 642.82 lux during full daylight periods

EFFECT PARAMETERS MEASURED: Immobilisation was recorded after 24 and 48 h test duration.

Reference substance (positive control):

no

Key result

Duration:

48 h

Dose descriptor:

EL50

Effect conc.:

> 1 000 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Remarks:

(WAF)

Basis for effect:

mobility

Details on results:

- Mortality of control: No mortality observed.
- Effect concentrations exceeding solubility of substance in test medium:

The quality of the dilution water used in culture and testing is monitored at weekly, monthly and semi-annual intervals. There are no known contaminants in the feed or water believed to be at levels high enough to interfere with this study based on contaminant analysis and/or historical data.

The TOC analysis indicates that the level of test substance in the WAF was at or near the level of organic carbon in the control. This result confirms the low solubility of the test substance in water and the unsuitability to use the measured concentrations for the determination of the effect value. Thus, the effect value is related to the nominal concentration.

Table 1: Measuremnt of the test concentration

Loading level [mg/L]	Total organic carbon [mg/L]*
	Day 0  “new”	Day 2 “old”
Control	19 ± 0.3	10 ± 0.8
1000	6.6 ± 0.3	16 ± 1.0

*: triplicate TOC measurements on each sample

Table 2: Immobilisation of Daphnia

Loading level [mg/L]	Percent immobilisation
	24 h	48 h
Control	0	0
62.5	0	5
125	0	0
250	0	0
500	0	0
1000	0	5

Reference 6

Endpoint:

short-term toxicity to aquatic invertebrates

Type of information:

read-across from supporting substance (structural analogue or surrogate)

Remarks:

5 substances are available for read across

Adequacy of study:

weight of evidence

Justification for type of information:

see section 13 for the attached justification

Reason / purpose for cross-reference:

read-across source

Reason / purpose for cross-reference:

read-across source

Reason / purpose for cross-reference:

read-across source

Reason / purpose for cross-reference:

read-across source

Reason / purpose for cross-reference:

read-across source

GLP compliance:

yes

Analytical monitoring:

no

Vehicle:

no

Key result

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

78 mg/L

Nominal / measured:

nominal

Basis for effect:

mobility

Remarks on result:

other: 95% confidence interval between 68 and 98 mg/L

Remarks:

Fatty acids, C5-10, esters with pentaerythritol (CAS 68424-31-7)

Key result

Duration:

48 h

Dose descriptor:

NOEC

Effect conc.:

32 mg/L

Nominal / measured:

nominal

Basis for effect:

mobility

Remarks on result:

other: Fatty acids, C5-10, esters with pentaerythritol (CAS 68424-31-7)

Key result

Duration:

48 h

Dose descriptor:

EL50

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Remarks on result:

other: 2,2-bis[(octanoyloxy)methyl]butyl decanoate (CAS 11138-60-6)

Key result

Duration:

48 h

Dose descriptor:

EL0

Effect conc.:

>= 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Remarks on result:

other: 2,2-bis[(octanoyloxy)methyl]butyl decanoate (CAS 11138-60-6)

Key result

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

> 1.3 mg/L

Nominal / measured:

meas. (geom. mean)

Conc. based on:

test mat.

Basis for effect:

mobility

Remarks on result:

other: Fatty acids, C5-9, tetraesters with pentaerythritol (CAS 67762-53-2)

Key result

Duration:

48 h

Dose descriptor:

EL50

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

other: test mat. WAF

Basis for effect:

mobility

Remarks on result:

other: Fatty acids, C5-9, tetraesters with pentaerythritol (CAS 67762-53-2)

Key result

Duration:

48 h

Dose descriptor:

NOEC

Effect conc.:

0.63 mg/L

Nominal / measured:

meas. (geom. mean)

Conc. based on:

test mat.

Basis for effect:

mobility

Remarks on result:

other: Fatty acids, C5-9, tetraesters with pentaerythritol (CAS 67762-53-2)

Key result

Duration:

48 h

Dose descriptor:

NOELR

Effect conc.:

46 other: % of a WSF prepared at a loading rate of 100 mg/L

Nominal / measured:

nominal

Conc. based on:

other: test mat. WAF

Basis for effect:

mobility

Remarks on result:

other: Fatty acids, C5-9, tetraesters with pentaerythritol (CAS 67762-53-2)

Key result

Duration:

48 h

Dose descriptor:

EL50

Effect conc.:

> 1 000 mg/L

Nominal / measured:

nominal

Conc. based on:

other: test mat./ (WAF)

Basis for effect:

mobility

Remarks on result:

other: Mixed esters of pentaerythritol, technical grade, with isooctanoic and C8/C10 fatty acids (CAS 189200-42-8)

Key result

Duration:

48 h

Dose descriptor:

EL50

Effect conc.:

> 1 072 mg/L

Nominal / measured:

nominal

Conc. based on:

other: test mat./WAF

Basis for effect:

mobility

Remarks on result:

other: Decanoic acid, mixed esters with heptanoic acid, octanoic acid, pentaerythritol and valeric acid (CAS 71010-76-9)

Validity criteria fulfilled:

not applicable

Remarks:

read across

Conclusions:

The substance, CAS 68424-31-7 ; EC 270-291-9, is analogous to the substances to be read across to, in terms of basic form, and the degree of substitution of functional groups is not considered to effect the proposed read across for the endpoint of short-term toxicity to aquatic invertebrates. Based on the available information to read across to, the substance is not expected to be toxic up to the limit of water solubility.

Description of key information

No toxic effects up to the limit of water solubility.

Additional information

In accordance to Regulation (EC) No. 1907/2006 Annex XI, 1.5 a read across to four structurally related category members decanoic acid, mixed esters with heptanoic acid, octanoic acid, pentaerythritol and valeric acid (CAS No. 71010-76-9), fatty acids, C5-9 tetraesters with pentaerythritol (CAS 67762-53-2), fatty acids, C8-10 (even numbered), di-and triesters with propylidynetrimethanol (CAS 11138-60-6) and fatty acids, C8-10 mixed esters with dipentaerythritol, isooctanoic acid, pentaerythritol and tripentaerythritol (CAS 189200-42-8) in order to fulfil the data requirements according to Annex VII-IX. This read-across is justified in detail in the overall summary (IUCLID chapter 6.1) and within the category justification in IUCLID Section 13. In this case of read-across, the best suited (highest degree of structural similarity, nearest physico-chemical properties) read-across substances were used for the assessment. The first two read-across substances have the same alcohol component, i.e. pentaerythritol, and are tetra-esters with a fatty acid chain length within the range of C5 – C10 comparable to the target substance.

The short-term toxicity of decanoic acid, mixed esters with heptanoic acid, octanoic acid, pentaerythritol and valeric acid (CAS No. 71010-76-9) to aquatic invertebrates was investigated according to OECD 202 using Daphnia magna as test organism under static condition (Blattenberger, 2006). The test solutions were prepared by adding the appropriate amount of test substance with subsequent stirring and sampling of the aqueous portions (WAFs) through the outlet at the bottom of the vessels. Loading rate WAFs of 62, 132, 251, 503 and 1072 mg/L were tested. No immobilisation was observed in any treatment and the control throughout the test period of 48 h. Hence, the 48 h-EL50 is determined to be > 1072 mg/L based on the nominal test concentration.

The short-term toxicity of fatty acids, C5-9 tetraesters with pentaerythritol (CAS 67762-35-2) to aquatic invertebrates was investigated in a study according to OECD 202 using Daphnia magna as test organism under static conditions (Migchielsen, 2012). The WAFs were prepared by adding the appropriate amount of test substance with subsequent stirring and sampling of the aqueous portions A nominal test concentration of 100 mg/L, corresponding to a measured concentration of 1.3 mg/L, was tested. No immobilisation was observed in the treatment throughout the test period of 48 h. Hence, the 48 h-EL50 is determined to be > 100 mg/L based on the nominal test concentration and > 1.3 mg/L based on the measured concentration. It can therefore be concluded that the test substance will not exhibit short-term effects to aquatic invertebrates up to the limit of water solubility.

In addition, two further read-across studies were available where the short term toxicity of fatty acids, C8-10 (even numbered), di-and triesters with propylidynetrimethanol (CAS-No. 11138-60-6), and fatty acids, C8-10 mixed esters with dipentaerythritol, isooctanoic acid, pentaerythritol and tripentaerythritol (CAS 189200-48-8) was investigated according to OECD 202. All measured results from 48 -hour toxicity studies indicate the test substance is of low toxicity toDaphnia magna. The study conducted with CAS 11138-60-6 (Häner, 2007) determined EL 50 (48h) > 100 mg/L (WAF loading rate) and Febbo (1995) observed an EL50 > 1000 mg/L, WAF loading rate, for CAS 189200-42-8.

These findings are supported by a short-term study with the target substance tetraesters from esterification of pentaerythritol with pentanoic, heptanoic and isononanoic acids (CAS 68424-30-6) rated as RL 4 due to insufficient documentation. In this study using Daphnia magna as test organism under static conditions (Windeatt, 1995) a 48 h-EC50 > 10 mg/L based on the nominal test concentration was determined. It can therefore be concluded that the test substance will not exhibit short-term effects to aquatic invertebrates up to the limit of water solubility.

Based on these results from structurally related read-across substances (in accordance to Regulation (EC) No 1907/2006 Annex XI, 1.5) which are characterized by a similar ecotoxicological profile, it can be concluded that pentaerythritol tetraesters of n-decanoic, n-heptanoic, n-octanoic and n-valeric acids (CAS 68424-31-7) will not exhibit short-term toxic effects to aquatic invertebrates up to the limit of water solubility.

As it can be seen in the data matrix of the category justification in section 13 and the overall summary, all reliable data in the category support the hazard assessment of each category member by showing a consistent pattern of results.